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Local and Systemic IKKε and NF-κB Signaling Associated with Sjögren's Syndrome Immunopathogenesis.

Chen W, Lin J, Cao H, Xu D, Xu B, Xu L, Yue L, Sun C, Wu G, Qian W - J Immunol Res (2015)

Bottom Line: In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative.Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity.These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China.

ABSTRACT
The activated NF-κB signaling pathway plays an important role in pathogenesis of primary Sjögren's syndrome (pSS). The inhibitor of κB (IκB) kinase (IKK) family such as IKKα, IKKβ, IKKγ, and IKKε, is required for this signaling. Our aim was to investigate the role of IKKα/β/γ/ε in patients with untreated pSS. In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative. And pIKKε positively related to expression of p-p65. Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity. Meanwhile, in peripheral blood mononuclear cells from pSS patients, pIKKε, total IKKε, pIKKα/β, and p-p65 were significantly increased by western blot, compared to healthy controls. However, there was no difference in IKKγ and IκBα between pSS patients and healthy individuals. These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

No MeSH data available.


Related in: MedlinePlus

pIKKε, pIκBα, and pNF-κB p65 were highly expressed in MSG from patients with pSS. Three pSS subgroups were classified by the grade of the inflammatory lesion (pSS-I: mild, pSS-II: intermediate, and pSS-III: severe lesions) by HE staining of MSG tissue (top row). The expression of pIKKε (second row, middle arrow), pIκBα (third row, large arrow), and pNF-κB p65 (bottom row, small arrow) was also detected in sections of MSG tissue by immunohistochemical (IHC) staining (brown), combined with counterstaining with Mayer's hematoxylin (blue). More severe infiltration grade was accompanied by higher expression of pIKKε, pIκBα, and pNF-κB p65. Representative examples are shown. Original magnification: ×200 for HE staining and ×400 for pIKKε, pIκBα, and pNF-κB p65 IHC. All data are representative of three independent experiments.
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fig1: pIKKε, pIκBα, and pNF-κB p65 were highly expressed in MSG from patients with pSS. Three pSS subgroups were classified by the grade of the inflammatory lesion (pSS-I: mild, pSS-II: intermediate, and pSS-III: severe lesions) by HE staining of MSG tissue (top row). The expression of pIKKε (second row, middle arrow), pIκBα (third row, large arrow), and pNF-κB p65 (bottom row, small arrow) was also detected in sections of MSG tissue by immunohistochemical (IHC) staining (brown), combined with counterstaining with Mayer's hematoxylin (blue). More severe infiltration grade was accompanied by higher expression of pIKKε, pIκBα, and pNF-κB p65. Representative examples are shown. Original magnification: ×200 for HE staining and ×400 for pIKKε, pIκBα, and pNF-κB p65 IHC. All data are representative of three independent experiments.

Mentions: First, we used immunohistochemical (IHC) staining to characterize the expression of proteins in the NF-κB pathway in minor salivary gland biopsies from patients with pSS and normal controls. The goal of this aspect of the study was to see whether NF-κB activation was related to severity of disease and to learn which cell types in the MSG tissue were sites of deregulated NF-κB activity. IHC analysis revealed that phosphorylated IKKε (pIKKε) was highly expressed in ductal epithelium and infiltrating mononuclear cells, but not in the acinar epithelium of MSG from patients with pSS (Figure 1). Phosphorylated IκBα was expressed in infiltrating mononuclear cells, ductal epithelium and acinar epithelium of MSG from patients with pSS (Figure 1). Total IKKε, total IKKα/β, and total IκBα were similarly expressed in ductal epithelium and mononuclear cells from pSS cases and in ductal epithelium from healthy controls. Total IκBα was also expressed at similarly high levels in acinar epithelium of MSG from both patients with pSS and healthy controls (Supplementary Figure S1, Table 2). However, no expression of pIKKα/β, total IKKγ, or pIKKγ was found in this tissue (Supplementary Figure S2, Table 2). Phosphorylation of the NF-κB family member p65 (p-p65), which indicates activation of NF-κB signaling, was observed in the infiltrating mononuclear cells and ductal epithelium in 93.9% (31/33) of patients with pSS, but in none of the healthy controls (Figure 1, Table 2).


Local and Systemic IKKε and NF-κB Signaling Associated with Sjögren's Syndrome Immunopathogenesis.

Chen W, Lin J, Cao H, Xu D, Xu B, Xu L, Yue L, Sun C, Wu G, Qian W - J Immunol Res (2015)

pIKKε, pIκBα, and pNF-κB p65 were highly expressed in MSG from patients with pSS. Three pSS subgroups were classified by the grade of the inflammatory lesion (pSS-I: mild, pSS-II: intermediate, and pSS-III: severe lesions) by HE staining of MSG tissue (top row). The expression of pIKKε (second row, middle arrow), pIκBα (third row, large arrow), and pNF-κB p65 (bottom row, small arrow) was also detected in sections of MSG tissue by immunohistochemical (IHC) staining (brown), combined with counterstaining with Mayer's hematoxylin (blue). More severe infiltration grade was accompanied by higher expression of pIKKε, pIκBα, and pNF-κB p65. Representative examples are shown. Original magnification: ×200 for HE staining and ×400 for pIKKε, pIκBα, and pNF-κB p65 IHC. All data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4563092&req=5

fig1: pIKKε, pIκBα, and pNF-κB p65 were highly expressed in MSG from patients with pSS. Three pSS subgroups were classified by the grade of the inflammatory lesion (pSS-I: mild, pSS-II: intermediate, and pSS-III: severe lesions) by HE staining of MSG tissue (top row). The expression of pIKKε (second row, middle arrow), pIκBα (third row, large arrow), and pNF-κB p65 (bottom row, small arrow) was also detected in sections of MSG tissue by immunohistochemical (IHC) staining (brown), combined with counterstaining with Mayer's hematoxylin (blue). More severe infiltration grade was accompanied by higher expression of pIKKε, pIκBα, and pNF-κB p65. Representative examples are shown. Original magnification: ×200 for HE staining and ×400 for pIKKε, pIκBα, and pNF-κB p65 IHC. All data are representative of three independent experiments.
Mentions: First, we used immunohistochemical (IHC) staining to characterize the expression of proteins in the NF-κB pathway in minor salivary gland biopsies from patients with pSS and normal controls. The goal of this aspect of the study was to see whether NF-κB activation was related to severity of disease and to learn which cell types in the MSG tissue were sites of deregulated NF-κB activity. IHC analysis revealed that phosphorylated IKKε (pIKKε) was highly expressed in ductal epithelium and infiltrating mononuclear cells, but not in the acinar epithelium of MSG from patients with pSS (Figure 1). Phosphorylated IκBα was expressed in infiltrating mononuclear cells, ductal epithelium and acinar epithelium of MSG from patients with pSS (Figure 1). Total IKKε, total IKKα/β, and total IκBα were similarly expressed in ductal epithelium and mononuclear cells from pSS cases and in ductal epithelium from healthy controls. Total IκBα was also expressed at similarly high levels in acinar epithelium of MSG from both patients with pSS and healthy controls (Supplementary Figure S1, Table 2). However, no expression of pIKKα/β, total IKKγ, or pIKKγ was found in this tissue (Supplementary Figure S2, Table 2). Phosphorylation of the NF-κB family member p65 (p-p65), which indicates activation of NF-κB signaling, was observed in the infiltrating mononuclear cells and ductal epithelium in 93.9% (31/33) of patients with pSS, but in none of the healthy controls (Figure 1, Table 2).

Bottom Line: In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative.Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity.These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China.

ABSTRACT
The activated NF-κB signaling pathway plays an important role in pathogenesis of primary Sjögren's syndrome (pSS). The inhibitor of κB (IκB) kinase (IKK) family such as IKKα, IKKβ, IKKγ, and IKKε, is required for this signaling. Our aim was to investigate the role of IKKα/β/γ/ε in patients with untreated pSS. In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative. And pIKKε positively related to expression of p-p65. Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity. Meanwhile, in peripheral blood mononuclear cells from pSS patients, pIKKε, total IKKε, pIKKα/β, and p-p65 were significantly increased by western blot, compared to healthy controls. However, there was no difference in IKKγ and IκBα between pSS patients and healthy individuals. These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

No MeSH data available.


Related in: MedlinePlus